CN107632032A - Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method - Google Patents
Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method Download PDFInfo
- Publication number
- CN107632032A CN107632032A CN201710808932.2A CN201710808932A CN107632032A CN 107632032 A CN107632032 A CN 107632032A CN 201710808932 A CN201710808932 A CN 201710808932A CN 107632032 A CN107632032 A CN 107632032A
- Authority
- CN
- China
- Prior art keywords
- mite
- male
- reproductive system
- female
- phyloseiulus nersimilis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Catching Or Destruction (AREA)
Abstract
The present invention relates to Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method, it is characterized in that the female mite of Phyloseiulus nersimilis, male mite in first being mated, soaked, be dehydrated by lactic acid again, dried, viscous platform, metal spraying are handled, the method that reproductive system is finally observed under ESEM.The present invention obtains Phyloseiulus nersimilis reproductive system first using very simple cleverly method, and the purpose that the trickle external structure of reproductive system is observed under ESEM is realized first, the acquisition of reproductive system can provide raw material to observe its structure under ESEM, and the result of ESEM is advantageous to the research to reproductive system.
Description
Technical field
The present invention relates to Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method, belong to Phyloseiulus nersimilis biology
Basic research field.
Background technology
Research shows that the Sex Determination period of Phyloseiulus nersimilis is occurred before ovum output, and even egg cell is just developed
It is front and rear.Therefore, it is a key point for understanding fully its reproduction mechanism that research is carried out to the growth course of Phyloseiulus nersimilis embryonated egg,
Study the growth course of embryonated egg, it is necessary to by its reproductive system（The development structure of phytoseiid mite embryonated egg）Dissection is out carried out
Research.
Electron microscopic observation is scanned to Phyloseiulus nersimilis reproductive system, can be from the knot of ultra micro reproductive system from horizontal
Structure form, this can be its clear and definite male mite sperm enter female mite body in after transport pathway and carried with the binding site of egg cell
For reference.
But because the too small comparison of individual of Phyloseiulus nersimilis is fragile, is full of in vivo by opaque body fluid, it is difficult to it
Reproductive system, which is accurately positioned, slightly accidentally may result in polypide crushes, and how available data dissects and such as without correlation again
What observes the report of its reproductive system, therefore how to obtain and be scanned electron microscopic observation Phyloseiulus nersimilis reproductive system, has
It is to be solved.
The content of the invention
, can be with using this method it is an object of the invention to provide Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method
Phyloseiulus nersimilis reproductive system is separately separated out and ultra micro observation is carried out under ESEM.
The present invention is the female mite of Phyloseiulus nersimilis, male mite in first being mated, then soaked by lactic acid, be dehydrated, dry,
Viscous platform, metal spraying processing, the method that reproductive system is finally observed under ESEM, concrete operation step are：
（1）The acquisition of male and female mite in mating：Under the microscope, out of Phyloseiulus nersimilis raising population, choose ovum and individually raise
To into progress male and female mite pairing after mite, needed according to different by the male and female individual of the different time in mating process together with raising
Device is positioned over ultra low temperature freezer together（-80℃）Or in liquid nitrogen, so as to obtain the male and female mite in different mating periods；
（2）Lactic acid soaks：After taking out the Phyloseiulus nersimilis male and female mite in mating, male and female mite is placed into slide with fine, soft fur pen
On, then with No. 0 insect needle and fine, soft fur pen male and female individual is separated, finally chosen male and female mite into the dactylethrae for filling lactic acid respectively
In, room temperature 3-4 days；
（3）Dehydration：Then take out female mite or male mite or be put into both in 70% alcohol together, then carry out serial dehydration, wine
The concentration of essence is followed successively by 80%, 90%, 100%, and specifically operation is first to be suctioned out upper strata alcohol with rubber head dropper, then adds again
Enter the alcohol of next gradient, so alcohol of the circulation until 100%；
（4）Dry：Taken female mite, male mite sample are placed on sample stage, are dried 2~3 hours using instrument is dried in vacuo；
（5）Viscous platform：Dried mite body backboard is gently taken off with No. 0 insect needle under the microscope, then with fine, soft fur pen by orange
The reproductive system of color takes out, and directly carries out viscous platform, and the internal structure if necessary to observe reproductive system needs light using insect needle
Light peels off reproductive system, is pressed lightly on insect needle, to ensure that polypide can be in close contact with conducting resinl；
（6）Metal spraying：It is gold-plated to mite whole body using ion sputtering instrument；
（7）Electron microscopic observation：It will be placed in mite sample stage in ESEM and be observed successively.
The present invention obtains Phyloseiulus nersimilis reproductive system first using very simple cleverly method, and realizes first
The purpose of the trickle external structure of reproductive system is observed under ESEM, the acquisition of reproductive system can be to observe its knot under ESEM
Structure provides raw material, and the result of ESEM is advantageous to the research to reproductive system.
Embodiment
Embodiment Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method
This method is the female mite of Phyloseiulus nersimilis, male mite in first being mated, then is soaked by lactic acid, be dehydrated, be dry, be viscous
Platform, metal spraying processing, the method that reproductive system is finally observed under ESEM, concrete operation step are：
（1）The acquisition of male and female mite in mating：Under the microscope, out of Phyloseiulus nersimilis raising population, choose ovum and individually raise
To into progress male and female mite pairing after mite, needed according to different by the male and female individual of the different time in mating process together with raising
Device is positioned over ultra low temperature freezer together（-80℃）Or in liquid nitrogen, so as to obtain the male and female mite in different mating periods；
（2）Lactic acid soaks：After taking out the Phyloseiulus nersimilis male and female mite in mating, male and female mite is placed into slide with fine, soft fur pen
On, then with No. 0 insect needle and fine, soft fur pen male and female individual is separated, finally chosen male and female mite into the dactylethrae for filling lactic acid respectively
In, room temperature 3-4 days；
（3）Dehydration：Then take out female mite or male mite or be put into both in 70% alcohol together, then carry out serial dehydration, wine
The concentration of essence is followed successively by 80%, 90%, 100%, and specifically operation is first to be suctioned out upper strata alcohol with rubber head dropper, then adds again
Enter the alcohol of next gradient, so alcohol of the circulation until 100%；
（4）Dry：Taken female mite, male mite sample are placed on sample stage, are dried 2~3 hours using instrument is dried in vacuo；
（5）Viscous platform：Dried mite body backboard is gently taken off with No. 0 insect needle under the microscope, then with fine, soft fur pen by orange
The reproductive system of color takes out, and directly carries out viscous platform, and the internal structure if necessary to observe reproductive system needs light using insect needle
Light peels off reproductive system, is pressed lightly on insect needle, to ensure that polypide can be in close contact with conducting resinl；
（6）Metal spraying：It is gold-plated to mite whole body using ion sputtering instrument；
（7）Electron microscopic observation：It will be placed in mite sample stage in ESEM and be observed successively.
Claims (1)
1. Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method, it is characterized in that the Phyloseiulus nersimilis in first being mated
Female mite, male mite, then soaked by lactic acid, be dehydrated, dry, viscous platform, metal spraying processing, finally observe reproductive system under ESEM
Method, concrete operation step is：
（1）The acquisition of male and female mite in mating：Under the microscope, out of Phyloseiulus nersimilis raising population, choose ovum and individually raise
To into progress male and female mite pairing after mite, needed according to different by the male and female individual of the different time in mating process together with raising
Device is positioned over ultra low temperature freezer together（-80℃）Or in liquid nitrogen, so as to obtain the male and female mite in different mating periods；
（2）Lactic acid soaks：After taking out the Phyloseiulus nersimilis male and female mite in mating, male and female mite is placed into slide with fine, soft fur pen
On, then with No. 0 insect needle and fine, soft fur pen male and female individual is separated, finally chosen male and female mite into the dactylethrae for filling lactic acid respectively
In, room temperature 3-4 days；
（3）Dehydration：Then take out female mite or male mite or be put into both in 70% alcohol together, then carry out serial dehydration, wine
The concentration of essence is followed successively by 80%, 90%, 100%, and specifically operation is first to be suctioned out upper strata alcohol with rubber head dropper, then adds again
Enter the alcohol of next gradient, so alcohol of the circulation until 100%；
（4）Dry：Taken female mite, male mite sample are placed on sample stage, are dried 2~3 hours using instrument is dried in vacuo；
（5）Viscous platform：Dried mite body backboard is gently taken off with No. 0 insect needle under the microscope, then with fine, soft fur pen by orange
The reproductive system of color takes out, and directly carries out viscous platform, and the internal structure if necessary to observe reproductive system needs light using insect needle
Light peels off reproductive system, is pressed lightly on insect needle, to ensure that polypide can be in close contact with conducting resinl；
（6）Metal spraying：It is gold-plated to mite whole body using ion sputtering instrument；
（7）Electron microscopic observation：It will be placed in mite sample stage in ESEM and be observed successively.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710808932.2A CN107632032A (en) | 2017-09-09 | 2017-09-09 | Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710808932.2A CN107632032A (en) | 2017-09-09 | 2017-09-09 | Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107632032A true CN107632032A (en) | 2018-01-26 |
Family
ID=61100494
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710808932.2A Pending CN107632032A (en) | 2017-09-09 | 2017-09-09 | Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107632032A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6058878A (en) * | 1998-06-17 | 2000-05-09 | California Polytechnic State University Foundation | Protozoan free colonies of lepidoptera |
| CN102607907A (en) * | 2012-02-24 | 2012-07-25 | 东北农业大学 | Paraffin section method for fern gametophytes |
| CN103868768A (en) * | 2014-02-14 | 2014-06-18 | 河南省农业科学院植物保护研究所 | Treatment method of scanning electron microscope samples of insect tentacles and appendages |
| CN104406834A (en) * | 2014-11-04 | 2015-03-11 | 中国水产科学研究院东海水产研究所 | Marine fish sperm staining method |
| CN105486554A (en) * | 2015-11-19 | 2016-04-13 | 新疆农业大学 | Formula and method for immobilizing tick sample for scanning electron microscopy |
-
2017
- 2017-09-09 CN CN201710808932.2A patent/CN107632032A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6058878A (en) * | 1998-06-17 | 2000-05-09 | California Polytechnic State University Foundation | Protozoan free colonies of lepidoptera |
| CN102607907A (en) * | 2012-02-24 | 2012-07-25 | 东北农业大学 | Paraffin section method for fern gametophytes |
| CN103868768A (en) * | 2014-02-14 | 2014-06-18 | 河南省农业科学院植物保护研究所 | Treatment method of scanning electron microscope samples of insect tentacles and appendages |
| CN104406834A (en) * | 2014-11-04 | 2015-03-11 | 中国水产科学研究院东海水产研究所 | Marine fish sperm staining method |
| CN105486554A (en) * | 2015-11-19 | 2016-04-13 | 新疆农业大学 | Formula and method for immobilizing tick sample for scanning electron microscopy |
Non-Patent Citations (2)
| Title |
|---|
| 吴桂华等: "《粉尘螨生殖系统形态学研究》", 《蠕形螨扫描电镜样本的制备方法》 * |
| 吴聪明: "《蠕形螨扫描电镜样本的制备方法》", 《动物学杂志》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Spohn et al. | Distribution of microbial-and root-derived phosphatase activities in the rhizosphere depending on P availability and C allocation–Coupling soil zymography with 14C imaging | |
| CN107632032A (en) | Phyloseiulus nersimilis reproductive system scanning electron microscopic observation method | |
| CN107389710A (en) | A kind of method for obtaining and small new neoseiulus californicus reproductive system being observed under ESEM | |
| CN107421793A (en) | The method that Predatory Mites reproductive system is observed under ESEM | |
| CN107655912A (en) | The acquisition of Tianjin river amblyseius as predatory mite reproductive system and its making observational technique under ESEM | |
| Popper et al. | The plant cell wall | |
| CN104818241A (en) | Skin tissue cell line of Epinephelus lanceolatus and establishing method thereof | |
| CN107589004A (en) | A kind of method for making and obtaining Tianjin river amblyseius as predatory mite reproductive system | |
| CN107494451A (en) | A kind of acquisition methods of small new neoseiulus californicus reproductive system | |
| CN107549119A (en) | The simple acquisition methods of Predatory Mites reproductive system | |
| CN107581157A (en) | A kind of method for directly obtaining Phyloseiulus nersimilis reproductive system | |
| CN114317440A (en) | Method for measuring forming ability of tumor organoid | |
| CN107655911A (en) | The acquisition of Tianjin river amblyseius as predatory mite reproductive system and its observational technique under transmission electron microscope | |
| CN107655904A (en) | Phyloseiulus nersimilis reproductive system transmission electron microscope observing method | |
| CN107632034A (en) | A kind of method for obtaining and small new neoseiulus californicus reproductive system being observed under transmission electron microscope | |
| CN107632033A (en) | A kind of method for obtaining and small new neoseiulus californicus spermatophore being observed under transmission electron microscope | |
| CN108267468A (en) | The method that Predatory Mites reproductive system is observed under transmission electron microscope | |
| CN107655913A (en) | The acquisition of Tianjin river amblyseius as predatory mite spermatophore and the method observed under ESEM | |
| CN107515314A (en) | A kind of method for obtaining and small new neoseiulus californicus spermatophore being observed under ESEM | |
| de Felipe et al. | Comparative study of two cryotechniques to elucidate real functional aspects of legume nodules development | |
| CN217241644U (en) | Plant culture device for soil-plant rhizosphere micro-area in-situ imaging | |
| CN107581156A (en) | A kind of acquisition methods of small new neoseiulus californicus spermatophore | |
| CN106350570B (en) | Real-time observation method for single cell of spore-forming bacteria | |
| CN107576682A (en) | The method that Predatory Mites spermatophore is observed under ESEM | |
| CN107568167A (en) | Phyloseiulus nersimilis hero mite spermatophore scanning electron microscopic observation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180126 |